Weidner J, Eigel A, Horst J, Köhnlein W
Institut für Strahlenbiologie, Westfälische Wilhelms-Universität, Münster, Germany.
Hum Mutat. 1994;4(1):55-6. doi: 10.1002/humu.1380040108.
This report describes a rapid convenient screening system with improved sensitivity to detect mutations in the cystic fibrosis transmembrane regulator (CFTR) gene based on nonisotopic SSCP analysis. Because conventional SSCP analysis is often hampered by poor yield of single-stranded DNA, we applied the well-established solid-phase technique in which streptavidin-coated magnetic beads are used to immobilize biotinylated polymerase chain reaction (PCR) products. High yield of single-stranded DNA can be eluted from the solid phase by denaturation and used for SSCP analysis. An additional advantage of this procedure is that the immobilized single strand is available without any further purification steps for solid-phase sequencing.
本报告描述了一种快速便捷的筛查系统，该系统基于非同位素单链构象多态性（SSCP）分析，对囊性纤维化跨膜传导调节因子（CFTR）基因中的突变检测具有更高的灵敏度。由于传统的SSCP分析常常受到单链DNA产量低的阻碍，我们应用了成熟的固相技术，其中使用链霉亲和素包被的磁珠来固定生物素化的聚合酶链反应（PCR）产物。通过变性可从固相中洗脱高产率的单链DNA，并用于SSCP分析。该方法的另一个优点是，固定的单链无需任何进一步的纯化步骤即可用于固相测序。
